1、BS EN 15742:2009ICS 65.120NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBRITISH STANDARDAnimal feeding stuffs Determination of OC-pesticides and PCBs by GC/ECDThis British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 July2009
2、 BSI 2009ISBN 978 0 580 60512 3Amendments/corrigenda issued since publicationDate CommentsBS EN 15742:2009National forewordThis British Standard is the UK implementation of EN 15742:2009. The UK participation in its preparation was entrusted to TechnicalCommittee AW/10, Animal feeding stuffs.A list
3、of organizations represented on this committee can be obtained onrequest to its secretary.This publication does not purport to include all the necessary provisionsof a contract. Users are responsible for its correct application. Compliance with a British Standard cannot confer immunityfrom legal obl
4、igations.BS EN 15742:2009EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN 15742February 2009ICS 65.120English VersionAnimal feeding stuffs - Determination of OC-pesticides andPCBs by GC/ECDAliments des animaux - Dtermination des pesticidesorganochlors (OC) et des polychlorobiphnyles (PCB) parGC/ECD
5、Futtermittel - Bestimmung der OC-Pestizide und PCBsmittels GC/ECD-VerfahrenThis European Standard was approved by CEN on 24 January 2009.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national
6、standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the CEN Management Centre or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any oth
7、er language made by translationunder the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, F
8、inland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOM
9、ITEE FR NORMUNGManagement Centre: Avenue Marnix 17, B-1000 Brussels 2009 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 15742:2009: EBS EN 15742:2009EN 15742:2009 (E) 2 Contents Page Foreword . 3 1 Scope 4 2 Normative references 4 3
10、 Terms and definitions . 4 4 Principle 5 5 Reagents and materials 5 6 Apparatus . 12 7 Sampling . 13 8 Preparation of test sample . 13 9 Procedure . 13 10 Calculation and expression of results. 15 11 Precision . 18 12 Test report 21 13 Important considerations . 21 Annex A (informative) Results of i
11、nterlaboratory tests . 22 Bibliography 23 BS EN 15742:2009EN 15742:2009 (E) 3 Foreword This document (EN 15742:2009) has been prepared by Technical Committee CEN/TC 327 “Animal feeding stuffs”, the secretariat of which is held by NEN. This European Standard shall be given the status of a national st
12、andard, either by publication of an identical text or by endorsement, at the latest by August 2009, and conflicting national standards shall be withdrawn at the latest by August 2009. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights
13、. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. This document has been prepared under a mandate given to CEN by the European Commission and the European Free Trade Association, and supports essential requirements of EC Directive(s). According to the
14、CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania
15、, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. BS EN 15742:2009EN 15742:2009 (E) 4 1 Scope This European Standard specifies a gas chromatographic method with electron capture detection (ECD) for the determin
16、ation of organochlorine pesticides (OCs) and polychlorinated biphenyls (PCBs) in animal feeding stuffs. The method is applicable to animal feeding stuffs with a water content up to about 20 wt% and oil/fatty samples containing residues of one or more of the following OCs, PCBs, toxaphene and some of
17、 their isomers and degradation products: Aldrin; Dieldrin; Chlorocamphene (Toxaphene); Chlordane (= sum of Chlordane isomers and Oxychlordane); DDT (= sum of isomers op-DDT, pp-DDT, pp-TDE (pp-DDD), and pp-DDE); Endosulfan (sum of -/-isomers and Endosulfan-sulphate); Endrin; Heptachlor (= sum of Hep
18、tachlor and -Heptachlorepoxide); Hexachlorobenzene (HCB); Hexachlorocyclohexane isomers -HCH (-BHC), -HCH (-BHC), -HCH (-BHC or lindane); PCB 28, 52, 101, 138, 153 and 180 (“Indicator PCBs”) and PCB 198, 209. The limit of quantification for the mentioned organochlorine pesticides and PCBs is 5 ng/g
19、in general. However, 10 ng/g applies for Heptachlor, Aldrin, Endrin, Dieldrin, and Endosulfan (-, - and sulphate). Individual laboratories are responsible to ensure that the equipment they used will achieve these limits of quantifications. 2 Normative references The following referenced documents ar
20、e indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 6498, Animal feeding stuffs Preparation of test samples 3 Terms and definitions Fo
21、r the purpose of this document, the following terms and definitions apply. 3.1 limit of detection smallest measured content, from which it is possible to deduce the presence of the analyte with reasonable statistical certainty BS EN 15742:2009EN 15742:2009 (E) 5 NOTE The limit of detection is numeri
22、cally equal to three times the standard deviation of the mean of blank determinations (n10). 3.2 limit of quantification lowest content of the analyte which can be measured with reasonable statistical certainty NOTE If both accuracy and precision are constant over a concentration range around the li
23、mit of detection, then the limit of quantification is numerically equal to 6 times the standard deviation of the mean of blank determinations (n10). 3.3 feed additives substances are feed additives when they comply with the definition of feed additives given in the Regulation 1831/2003 4 Principle A
24、 test portion of animal feeding stuff is fortified with internal standard (PCB 198), and is extracted with ethylacetate. The extract is concentrated and subsequently purified by: Gel permeation chromatography (GPC), with cyclohexane/ethylacetate as eluting solvent chromatography on partially deactiv
25、ated silica gel. The collected fraction containing the compounds of interest is concentrated and re-dissolved in a solution containing another internal standard (PCB 209) as a reference standard. After cleanup the analytes are measured using GC-ECD. Indentification is done on the basis of comparing
26、retention times on capillary columns of different polarity. Quantification is done using the internal standard method. 5 Reagents and materials Use only reagents of recognized analytical grade and with a purity suitable for OC and PCB residue analysis. Check the purity of the reagents by performing
27、a blank test under the same conditions as used in the method. The chromatogram should not show any interfering impurity at the retention time of compounds of interest. WARNING The use of this European Standard can involve hazardous materials, operations and equipment. This standard does not purport
28、to address all the safety problems associated with its use. It is the responsibility of the user of this European Standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. 5.1 Cyclohexane 5.2 Ethylacetate 5.3 Hexane 5.4 Dich
29、loromethane 5.5 Iso-octane 5.6 Toluene BS EN 15742:2009EN 15742:2009 (E) 6 5.7 Hexane/toluene = 3+7, parts by volume Mix 30 ml of hexan (5.3) with 70 ml of toluene (5.6) thoroughly. Store at room temperature in a tightly closed glass bottle. 5.8 Sodium Sulphate, anhydrous Heated at 160C during at le
30、ast 24 h. 5.9 Ethylacetate/Cyclohexane = 1+1, parts by volume Mix 500 ml of ethylacetate (5.2) with 500 ml of cyclohexane (5.1) thoroughly. Store at room temperature in a tightly closed glass bottle. 5.10 Silica gel, deactivated with 3,5% water Heat silica gel 60 (63m to 200m = 70 mesh to 230 mesh),
31、 at 130C for at least 5 h, allow to cool in a desiccator, and store in a tightly stopped container in the desiccator. To 96,5 g dried silica gel in a 300 ml Erlenmeyer flask with a ground joint, add 3,5 ml water dropwise from a burette, with continuous swirling. Immediately stopper the flask with a
32、ground stopper and shake vigorously for 5 min until all lumps have disappeared. Next shake for 2 h on a mechanical shaker, and then store in a tightly stoppered container. Deactivated silica gel is tenable during approximately 2 weeks if carefully stored. 5.11 Internal standard (PCB 198) 5.12 Intern
33、al Standard (PCB 209) 5.13 OC-pesticide reference standards Each with a purity not less than 99%. Aldrin (1R,4S,4aS,5S,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5,8-dimethanonaphthalene) CAS Number: 309-00-2 Dieldrin (1R,4S,4aS,5R,6R,7S,8S,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,
34、7,8,8a-octahydro-6,7epoxy-1,4:5,8-dimethanonaphthalene) CAS Number: 60-57-1 Chlordane (1,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-ethano-1H-indene); and isomer CAS Numbers: 5103-71-9 and 5103-74-2 Oxychlordane (4,7-Methanoindan, 1,2,4,5,6,7,8,8-octachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-,
35、 exo,endo-) CAS Number: 27304-13-8 op-DDT o,p-(1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) CAS Number: 789-02-6 BS EN 15742:2009EN 15742:2009 (E) 7 pp-DDT p,p-(1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) CAS Number: 50-29-3 pp-TDE (p,p-DDD) p,p-1,1-dichloro-2,2-bis(4-chlorophenyl) ethane CAS
36、Number: 72-54-8 pp-DDE p,p-(1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene) CAS Number: 72-55-9 Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin 3-oxide) two stereoisomers, , (I), CAS Number: 959-98-8 and , (II), CAS Number: 33213-65-9. Endosulfan-sulph
37、ate; CAS Number: 1031-07-8 Endrin (1R,4S,4aS,5S,6S,7R,8R,8aR)-1,2,3,4,10,10-hexachloro-1,4,4a,5,6,7,8,8a-octahydro-6,7-epoxy-1,4:5,8-dimethanonaphthalene CAS Number: 72-20-8 Heptachlor (1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene) CAS Number: 76-44-8 -Heptachlorepoxide (1,4,5,6,
38、7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene(exo) CAS Number: 1024-57-3 HCB (hexachlorobenzene) CAS Number: 118-74-1 -HCH (-BHC) (-1,2,3,4,5,6-hexachlorocyclohexane) CAS Number: 319-84-6 -HCH (-BHC) (-1,2,3,4,5,6-hexachlorocyclohexane) CAS Number: 319-85-7 -HCH (-BHC; lindane) (1,2,3,4,5
39、,6-hexachlorocyclohexane) CAS Number: 58-89-9 Or a Certified Mixture at a concentration of 10 g/ml. 5.14 PCBs reference standards Each with a purity not less than 99%. PCB 28 (2,4,4 trichlorobiphenyl) ; CAS Number: 7012-37-5 PCB 52 (2,2,5,5 tetrachlorobiphenyl) ; CAS Number: 35693-99-3 BS EN 15742:2
40、009EN 15742:2009 (E) 8 PCB 101 (2,2,4,5,5 pentachlorobiphenyl) ; CAS Number: 37680-73-2 PCB 138 (2,2,3,4,4,5 hexachlorobiphenyl) ; CAS Number: 35065-28-2 PCB 153 (2,2,4,4,5,5 hexachlorobiphenyl) ; CAS Number: 35065-27-1 PCB 180 (2,2,3,4,4,5,5 heptachlorobiphenyl) ; CAS Number: 35065-29-3 Or a Certif
41、ied Mixture at a concentration of 10 g/ml. 5.15 Chlorocamphene (Toxaphene) Technical mixture. 5.16 Stock solutions, 100 g/ml Weigh 5 -10 mg ( 0,01 mg) of each compound (5.11, 5.12, 5.13, 5.14 and 5.15) in seperate brown medicine glass bottles of 100 ml and add iso-octane (5.5) to achieve a concentra
42、tion of 100 g/ml. Store the solutions in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. Dissolve -HCH in 10 ml toluene (5.6), to achieve complete solvability and dilute further with iso-octane (5.5) to achieve
43、a concentration of 100 g/ml. 5.17 Mixed stock solutions 5.17.1 Mixed stock solution OC (without Endosulfan and Toxaphene) Pipet of each OC-stock solution (5.16) the indicated volume (Table 1) in a volumetric flask of 100 ml. Fill up to 100 ml with iso-octane (5.5) and mix. The achieved concentration
44、 is given in Table 1. Transport this solution to a brown medicine glass bottle of 100 ml and store it in a refrigerator at 4C ( 3C). The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. 5.17.2 Mixed stock solution Endosulfan Pipet of each Endos
45、ulfan-stock solution (5.16) the indicated volume (Table 1) in a volumetric flask of 100 ml. Fill up to 100 ml with iso-octane (5.5) and mix. The achieved concentration is given in Table 1. Transport this solution to a brown medicine glass bottle of 100 ml and store it in a refrigerator at 4C ( 3C).
46、The solution is tenable under these conditions during at least 5 years if the weight is carefully controled. BS EN 15742:2009EN 15742:2009 (E) 9 Table 1 Concentration of OCs in Mixed stock solution (5.17) and mixed standard solution (5.18) Compound Pipet volume (ml) Mixed stock solution (5.17 1 Aisi
47、s the area internal standard PCB 198; Arsis the area of PCB 209; Qisis the amount of internal standard PCB 198 ng/ml; Qrsis the amount of PCB 209 ng/ml; Qxis the amount of component of interest ng/ml. Consequently the averaged relative response factor is calculated: (n)RRFx1=RRF i=i(n)524(3) where n
48、 is the component of interest; BS EN 15742:2009EN 15742:2009 (E) 17 i is the calibration level (2 thru 5). Consequently the averaged relative response factor is calculated for internal standard (PCB 198): (m)RRFx1=RRF i=i(m)524(4) where m is the internal standard (PCB 198); i is the calibration leve
49、l (2 thru 5); Calculation concentration component of interest The concentration component of interest is calculated by: RRFxWxAQxA=C(n)isisxx(5) where Cxis the concentration component of interest in ng/g; Ax is the area of component of interest in the sample extract; Ais is the area of used internal standard (PCB 198) in the sample extract; Qisis the amount of internal standard (PCB 198) ng/ml; W is the weight of injected sample amount equivalent g/ml
